• 제목/요약/키워드: Marker-less

검색결과 235건 처리시간 0.028초

HPLC를 이용한 노루오줌 추출물의 Avicularin 및 Quercitrin 분석법 개발 (Analytical Method Development of Avicularin and Quercitrin in Astilbe chinensis Extract Using HPLC)

  • 권진관;정연우;서찬곤;홍성수;신현탁;정수영;최정준;김진규
    • 대한화장품학회지
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    • 제45권4호
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    • pp.333-340
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    • 2019
  • 본 연구는 노루오줌(Astilbe chinensis) 추출물을 기능성 화장품소재로 개발하기 위해 avicularin, quercitrin을 지표성분으로 선정하고, 품질관리를 위해 high performance liquid chromatography (HPLC)를 이용하여 분석법을 개발하였다. 분석용 칼럼은 Capcell Pak C18 MGII을, 이동상은 0.05% (v/v) trifluoroacetic acid (TFA)와 아세토니트릴을 사용하여 컬럼 온도는 30℃ 에서 유속 1.0 mL/min 로 검출파장은 254 nm에서 검출하였다. International conference on harmonization (ICH) 가이드라인(version 4, 2005)을 근거로 하여 특이성, 직선성, 정밀성, 정확성, 검출한계 및 정량한계를 분석하여 분석방법을 검증하였다. 분석결과, 검출한계 및 정량한계는 avicularin 0.094 및 0.285 mg/mL, quercitrin 0.031 및 0.095 mg/mL로 나타났으며, 검량곡선은 상관계수 값이 avicularin 0.99990, quercitrin 0.99994로 양호한 직선성을 보였고 정밀성 분석결과도 avicularin 0.59%, quercitrin 0.63% 이하로 확인되었다. 또한, 회수율은 avicularin 100.97 ~ 101.77%, quercitrin 100.18 ~ 100.32% 범위로 정확성이 있음을 알 수 있다. 따라서, 본 분석법은 노루오줌 추출물 지표성분의 분석법으로 적합한 시험법임이 검증되었다.

Prognostic Significance of 14-3-3γ Overexpression in Advanced Non-Small Cell Lung Cancer

  • Raungrut, Pritsana;Wongkotsila, Anusara;Lirdprapamongkol, Kriengsak;Svasti, Jisnuson;Geater, Sarayut Lucien;Phukaoloun, Monlika;Suwiwat, Supaporn;Thongsuksai, Paramee
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권8호
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    • pp.3513-3518
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    • 2014
  • The 14-3-3 protein has been shown to be involved in the cancer process. However, there is no understanding of the relationship between 14-3-$3{\gamma}$ (14-3-3 gamma) expression and prognosis in advanced non-small cell lung cancer. In this study, we therefore investigated the association between protein levels by immunohistochemistry and clinicopathological features of advanced NSCLC patients. Survival curves were estimated using the Kaplan-Meier method and tested by log-rank. Multivariate analysis was conducted with the Cox's regression model to determine independence of factors. p values less than 0.05 were considered significant. A total 153 patients were studied, with 54.3% being stage III and 45.8% stage IV. Fifty-one cases (33.3%) were squamous cell carcinomas, and 98 cases (64.1%) were adenocarcinomas. High 14-3-$3{\gamma}$ expression was seen in 59.5% and significantly correlated with lymph node metastasis (p=0.010) and distant metastasis (p=0.017). On Kaplan-Meier analysis, high 14-3-$3{\gamma}$ expression was associated with poorer survival with a marginal trend toward significance (p=0.055). On multivariate analysis, age, treatment, and 14-3-$3{\gamma}$ expression proved to be independent prognostic parameters. In vitro experiments indicated that 14-3-$3{\gamma}$ overexpression also played a potential role in cancer invasion. In conclusion, our data suggest that 14-3-$3{\gamma}$ overexpression is associated with invasion and a poor prognosis. Therefore, 14-3-$3{\gamma}$ may be a potential prognostic marker of advanced non-small cell lung cancer.

Prognostic Significance of Interactions Between ER Alpha and ER Beta and Lymph Node Status in Breast Cancer Cases

  • Han, Shu-Jing;Guo, Qing-Qing;Wang, Ting;Wang, You-Xin;Zhang, Yu-Xiang;Liu, Fen;Luo, Yan-Xia;Zhang, Jie;Wang, You-Li;Yan, Yu-Xiang;Peng, Xiao-Xia;Ling, Rui;He, Yan
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권10호
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    • pp.6081-6084
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    • 2013
  • Objective: Both estrogen receptors, ER alpha ($ER{\alpha}$) and ER beta ($ER{\beta}$), are expressed in 50-70% of breast cancer cases. The role of $ER{\alpha}$ as a prognostic marker in breast cancer has been well established as its expression is negative correlated with tumor size and lymph node metastasis. $ER{\beta}$ is also a favorable prognostic predictor although this is less well documented than for $ER{\alpha}$. Materials and Methods: To explore whether ERs independently or together might influence clinical outcome in breast cancer, the correlation between the ERs with the clinicopathological features was analyzed in 84 patients. Results: $ER{\alpha}$ expression negatively correlated with tumor stage (r=-0.246, p=0.028) and tended to be negatively correlated with lymph node status (r=-0.156, p=0.168) and tumor size (r=-0.246, p=0.099). Also, $ER{\beta}$ was negatively correlated with nodal status (r=-0.243, p=0.028), as was coexpression of $ER{\alpha}$ and $ER{\beta}$ (p=0.043, OR=0.194, 95% CI= 0.040-0.953). Conclusion: Coexpression of ERs might serve as an indicator of good prognosis in breast cancer patients.

Screening for Lynch Syndrome in Young Colorectal Cancer Patients from Saudi Arabia Using Microsatellite Instability as the Initial Test

  • Alqahtani, Masood;Grieu, Fabienne;Carrello, Amerigo;Amanuel, Benhur;Mashour, Miral;Alattas, Rabab;Al-Saleh, Khalid;Alsheikh, Abdulmalik;Alqahtani, Sarah;Iacopetta, Barry
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권4호
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    • pp.1917-1923
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    • 2016
  • Background: Lynch Syndrome (LS) is a familial cancer condition caused by germline mutations in DNA mismatch repair genes. Individuals with LS have a greatly increased risk of developing colorectal cancer (CRC) and it is therefore important to identify mutation carriers so they can undergo regular surveillance. Tumor DNA from LS patients characteristically shows microsatellite instability (MSI). Our aim here was to screen young CRC patients for MSI as a first step in the identification of unrecognized cases of LS in the Saudi population. Materials and Methods: Archival tumor tissue was obtained from 284 CRC patients treated at 4 institutes in Dammam and Riyadh between 2006 and 2015 and aged less than 60 years at diagnosis. MSI screening was performed using the BAT-26 microsatellite marker and positive cases confirmed using the pentaplex MSI analysis system. Positive cases were screened for BRAF mutations to exclude sporadic CRC and were evaluated for loss of expression of 4 DNA mismatch repair proteins using immunohistochemistry. Results: MSI was found in 33/284 (11.6%) cases, of which only one showed a BRAF mutation. Saudi MSI cases showed similar instability in the BAT-26 and BAT-25 markers to Australian MSI cases, but significantly lower frequencies of instability in 3 other microsatellite markers. Conclusions: MSI screening of young Saudi CRC patients reveals that approximately 1 in 9 are candidates for LS. Patients with MSI are strongly recommended to undergo genetic counselling and germline mutation testing for LS. Other affected family members can then be identified and offered regular surveillance for early detection of LS-associated cancers.

cDNA Cloning and Polymorphism of the Porcine Carbonic Anhydrase III (CA3) Gene

  • Wu, J.;Deng, Changyan;Xiong, Y.Z.;Zhou, D.H.;Lei, M.G.;Zuo, B.;Li, F.E.;Wang, J.
    • Asian-Australasian Journal of Animal Sciences
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    • 제19권3호
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    • pp.324-328
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    • 2006
  • Carbonic anhydrase III (CA3) is a member of a multigene family that encode carbonic anhydrase isozymes. In this study, a complete coding sequence of the pig CA3 gene which encodes a 260 amino-acid protein was determined. The amino acid comparison showed high sequence similarities with previously identified human (86.5%) CA3 gene and mouse (91.5%) Car3 gene. The partial genomic DNA sequences were also investigated. The length of intron 1 was 727 bp. Comparative sequencing of three pig breeds revealed that there was a T${\rightarrow}$C substitution at position 363 within intron 1. The substitution was situated within a NcoI recognition site and was developed as a PCR-restriction fragment length polymorphism (RFLP) marker for further use in population variation investigations and association analysis. Two alleles (A and B) were identified, and 617 bp fragments were observed for the AA genotype and 236 bp and 381 bp fragments for the BB genotype. The polymorphism of CA3 was detected in 8 pig breeds. Allele B was predominant in the Western pig breeds. In addition, association studies of the CA3 polymorphism with carcass traits in 140 $Yorkshire{\times}Meishan$ $F_2$ offspring showed that the NcoI PCR- RFLP genotype may be associated with variation in several carcass traits of interest for pig breeding. Allele B was associated with increases in lean meat percentage, loin eye height and loin eye area. Statistically significant association with backfat thickness was also found; pigs with the AB genotype had much less backfat thickness than AA or BB genotypes.

Detection of Recombinant Marker DNA in Genetically Modified Glyphosate- Tolerant Soybean and Use in Environmental Risk Assessment

  • Kim, Young-Tae;Park, Byoung-Keun;Hwang, Eui-Il;Yim, Nam-Hui;Lee, Sang-Han;Kim, Sung-Uk
    • Journal of Microbiology and Biotechnology
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    • 제14권2호
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    • pp.390-394
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    • 2004
  • The genetically modified glyphosate-tolerant soybean contains the following introduced DNA sequences: the EPSPS (5-enol-pyruvylshikimate-3-phosphate synthase) gene from Agrobacterium sp. strain CP4, the 35S promoter from the cauliflower mosaic virus, and the NOS terminator from Agrobacterium tumefaciens. In the present study, detection of these introduced DNAs was performed by amplification using the polymerase chain reaction (PCR). A multiplex PCR method was also applied to prevent false positive results. When primers for 35S promoter, nos3', CTP(chloroplast transit peptide), and CP4 EPSPS (EPSPS from Agrobacterium sp. CP4) were used, positive results were obtained in PCR reactions using DNA from genetically modified glyphosate-tolerant soybeans. There were no false positive results when using DNA from non-genetically modified soybeans. The CP4 EPSPS gene was detected when less than 125 pg glyphosate-tolerant soybean DNA was amplified. Lectin Lel and psb A were amplified from both non-genetically modified and genetically modified glyphosate-tolerant soybean DNA. Multiplex PCR was performed using different primer sets for actin Sacl, 35S promoter and CP4 EPSPS. The actin gene was detectable in both non-genetically modified and glyphosate-tolerant soybeans as a constant endogenous gene. Target DNAs for the 35S promoter, and CP4 EPSPS were detected in samples containing 0.01-0.1% glyphosate-tolerant soybean, although there were variations depending on primers by multiplex PCR. Soybean seeds from five plants of non-genetically modified soybean were co-cultivated for six months with those of genetically modified soybean, and they were analyzed by PCR. As a result, they were not positive for 35S promoter, nos3' or CP4 EPSPS. Therefore, these results suggest there was no natural crossing of genes between glyphosate-tolerant and non-genetically modified soybean during co-cultivation, which indicates that gene transfer between these plants is unlikely to occur in nature.

경부 재발 갑상선 유두암 환자에서 혈청 갑상선글로불린의 임상적 의의 (Clinical Implication of Serum Thyroglobulin in Recurred Papillary Thyroid Cancer at Neck Nodes)

  • 이하나;한명월;이호준;노종렬;남순열;김상윤;최승호
    • 대한두경부종양학회지
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    • 제27권1호
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    • pp.42-46
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    • 2011
  • Background and Objectives : Serum stimulated thyroglobulin(stim Tg) was well-known for useful marker in detecting of recurrent or persistent papillary thyroid cancer after total thyroidectomy. Serum stim Tg level may be possibly related with recurrent tumor volume, but rarely studied. The purpose of this study was to examine the relationship between preoperative serum stim Tg level and recurrent tumor burden and to find additional clinical usefulness of stim Tg more than to detect a recurrence. Material and Methods : From January 2000 to December 2009, 40 patients who were operated due to neck recurrence of papillary thyroid cancer after total thyroidectomy were enrolled. All patients had preoperative stim Tg. We compared the clinical correlation of stim Tg and other variables to influence the preoperative stim Tg levels. Results : Preoperative stim Tg levels weren't correlated with site of recurrence, number of metastasis, maximal size, and presence of extra-capsular spread. But considerable increase of stim Tg more than 50ng/mL was identified in recurrence of lateral neck. Patients who have higher stim Tg level after surgery tend to be have higher preoperative stim Tg level. Conclusion : stim Tg was not elevated in 7.5% of recurrent PTC patients. Thus, other diagnostic modalities such as US may be important for these patients. If preoperative stim Tg was more than 50ng/mL, it may suggest recurrence in lateral neck and have less possibility to achieve postoperative biochemical remission.

사람 폐 섬유아 세포에서 Brunfelsia grandiflora 에탄올 추출물이 Autophagy에 미치는 영향 (The Effect of Brunfelsia grandiflora Ethanol Extract on the Induction of Autophagy in Human Lung Fibroblasts)

  • 남향;김문무
    • 생명과학회지
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    • 제24권8호
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    • pp.837-842
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    • 2014
  • 이 연구의 목적은 사람 폐 섬유아세포인 IMR 90 에서 Brunfelsia grandiflora 에탄올 추출물(BGEE)이 SIRT1 및 p53 활성화를 통해 autophagy의 유도에 대한 효과를 조사한 것이다. BGEE는 $5{\mu}g/ml$ 이상의 농도에서 IMR 90 세포에서 세포독성을 나타내었다. 본 연구에서 처음으로 BGEE가 autophagy를 유도 하는 것이 발견되었다. 또한, BGEE는 $2.5{\mu}g/ml$ 이하에서 Beclin-1 및 $5{\mu}g/ml$ 이상에서 Atg7 의 활성화가 autophagy의 유도에 관여함을 확인하였다. 더욱이 BGEE는 autophagy와 관련된 단백질 발현을 조절하였는데 p53 및 p-p53 단백질 발현이 세포독성이 없는 농도의 BGEE존재하에서 감소되었다. 하였다. 반면에, SIRT1의 발현수준은 세포독성이 없는 농도의 BGEE로 처리된 IMR 90 세포에서 증가되었다. 더욱이 BGEE로 처리된 사람 페 섬유아세포에서 노화 마커의 지표인 SA-${\beta}$-gal staning이 감소되는 것이 관찰되었다. 이상의 발견들은 BGEE는 사람 폐 섬유아세포에서 p53 및 SIRT1의 조절을 통하여 autophagy 및 항노화 유발을 촉진 시키는 것을 시사하고 있다.

소나무 천연갱신림내 성목과 치수의 유전변이 비교 (Comparison of Genetic Variation between Pre-practice Mature Trees and Post-practice One-year Old Seedlings in Pinus densiflora Natural Regeneration Stands)

  • 안지영;이제완;이석우;백승훈;임효인;김현섭
    • 한국산림과학회지
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    • 제104권4호
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    • pp.543-548
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    • 2015
  • 단목모수, 군상모수, 군상개벌, 대상개벌 등의 천연갱신 방법이 적용된 소나무림의 nSSR 유전변이를 조사하였다. 작업(벌채) 전 성목과 작업 후 임내에서 자연 발생한 1년생 치수들의 유전다양성을 비교한 결과 큰 차이가 없는 것으로 나타났으며(성목: A=13.4; $A_e$=4.3; $H_o$=0.596; $H_e$=0.598, 치수: A=13.6; $A_e$=4.3; $H_o$=0.571; $H_e$=0.597), 각각의 작업종에서도 통계적으로 유의한 차이를 발견할 수 없었다. 성목과 치수의 유전분화 정도는 매우 낮은 것으로 나타났으며($F_{ST}$=0.002), 각 작업종별 유전분화 정도 역시 낮은 값($$F_{ST}{\leq_-}0.01$$)을 보여 작업 전, 후 임분의 유전구조 변화는 크지 않은 것으로 추정되었다. 결과적으로 본 연구가 수행된 소나무 임분의 경우 유전다양성과 유전적 조성 변화에 미치는 천연갱신 작업종의 효과가 두드러지지 않은 것으로 나타났다. 그러나, 천연갱신 작업종이 유전변화에 미치는 영향을 보다 정확하게 평가하기 위해서는 작업 후 남겨진 개체(모수)의 교배로부터 발생한 치수들의 시계열적 유전변이와 유전구조 변화를 지속적으로 추정할 필요가 있는 것으로 판단된다.

Construction of Various Copy Number Plasmid Vectors and Their Utility for Genome Sequencing

  • Yang, Tae-Jin;Yu, Yeisoo;Frisch, David A.;Lee, Seunghee;Kim, Hye-Ran;Kwon, Soo-Jin;Park, Beom-Suk;Wing, Rod A.
    • Genomics & Informatics
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    • 제2권4호
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    • pp.174-179
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    • 2004
  • We developed various plasmid cloning vectors that are useful in the construction of genomic and shotgun libraries. Two medium copy vectors, pCUGlblu21 (pCb21) and pAGlblu21 (pAb21), which are resistant to kanamycin ($Km^R$) and chloramphenicol ($Cam^R$), respectively, are useful for cloning DNA inserts ranging from 5kb to 15kb. Two high copy vectors, pCUGlblu31 (pCb31) and pAGlblu31 (pAb31), containing $Km^R$ and $Cam^R$, respectively, are useful for DNA inserts less than 5kb. These vectors are well adapted for large-scale genome sequencing projects by providing choice of copy number and selectable marker. The small vector size is another advantage of these vectors. All vectors contain lacZa including multicloning sites that originated from pBluscriptllsk- for easy cloning and sequencing. Two medium copy vectors contain unique and rare cutting Swal (ATTTAAAT) restriction enzyme sites for easy determination of insert size. We developed two combined vectors, pC21A31 and pC31A21, which are combinations of (pCb21 + pAb31) and (pCb31 + pAb21), respectively. These two vectors provide four choices of vectors such as $Km^R$ and medium, $Cam^R$ and high, $Cam^R$ and medium, and $Km^R$ and high copy vectors by restriction enzyme cutting, dephosphorylation, and gel purification. These vectors were successfully applied to high throughput shotgun sequencing of rice, tomato, and brassica BAC clones. With an example of extremely biased hydro sheared 3 kb shotgun library of a tomato BAC clone, which is originated from cytogenetically defined peri-centromeric region, we suggest the utility of an additional 10 kb library for sequence assembly of the difficult-to-assemble BAC clone.